development of a sav tissue culture protocol for …...development of a sav tissue culture protocol...
TRANSCRIPT
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Development of a SAV Tissue Culture Protocol for
Restoration of Marine Habitats
Kayti Tigani
May 18, 2007Center for the Inland Bays
Scientific & Technical Advisory Committee
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3 CommonSAV Restoration Pitfalls
• Lack of plant material supplies
• Local plant population
• Plants with specific characteristics for the restoration site
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Plant Tissue Culture 101
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Totipotent
Every plant cell has the potential to divide and produce all the cells to build a new organism
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Plant Tissue Culture 101
Explant
DonorPlant
Culture DishMedia
Micropropagation
Callus Induction
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Plant Tissue Culture 101
Differentiation
Propagules
Micropropagation
DifferentiationCallus Induction
Callus
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Seagrass Tissue Culture & Restoration
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Plant Material Source
• Less collection & more material• Uses the local genotype• Guaranteed availability
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Selective Cultivars
These plants could…
Survive more turbidity?
Withstand nutrient loading?
Higher root:shoot ratio?
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Band-Aid CropSeedling alone Band-Aid Crop
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Genotype Banking
Dredging
Indian River Inlet Bridge Construction
www.faust-corp.com
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Tissue Culture & Research
• In vitro studies of uptake and production
• Microbial community dynamics
• Diagnosis of future events
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Development of a Tissue Culture Protocol
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Halophyte Biotechnology Center
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The NOAA/UNH Cooperative Institute for Coastal and Estuarine
Environmental Technology (CICEET)
CICEET
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Field Collection
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Back To The Lab…
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A Two-Tract Approach
Callus InductionMicropropagation
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2-Phase Media
Liquid topmedia layer
Solid bottommedia layer
2-Phase media for micropropagation / shoot proliferation
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Sterilization Technique
• Ethanol• 10% Betadine • 0.2 mg/L erythromycin
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Sterilization Technique
The Result:
Contaminated culture Sterile culture
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Common Bacterial Association
16S rRNA genesequence data andfatty acid analysis
revealedThalassospira spp.
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Current/Future Strategies
• New hormone combinations for micropropagation
• Hormone pulse method
• Organic media nutrients
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End-User Advisor Roll
Ben Anderson DNREC
Otto Bundy Environmental Plant Resources, Inc.
Beau Williams Seagrass Recovery, Inc.
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Development of a SAV Tissue Culture Protocol for
Restoration of Marine Habitats
Kayti TiganiHalophyte Biotechnology Center